New metabolically stable fatty acid amide ligands of cannabinoid receptors: synthesis and receptor affinity studies

We investigated the structure-activity relationships for the interactions of fatty acid amide analogs of the endocannabinoid anandamide with human recombinant cannabinoid receptors. Thirty-five novel fatty acid amides were synthesized using five different types of acyl chains and 11 different aromatic amine 'heads.' Although none of the new compounds was a more potent ligand than anandamide, we identified three amine groups capable of improving the metabolic stability of arachidonoylamides and their CBI/CB2 selectivity ratio to over 20-fold, and several aromatic amines capable of improving the affinity of short chain or monosaturated fatty acids for cannabinoid CBI receptors. For the first time a tertiary amide of arachidonic acid was found to possess moderate affinity (K, = 300 nM) for cannabinoid CBI, but not CB2, receptors

Structure-Activity Relationships and Computational Studies on PDE4 inhibitors

Supplement 1, Sci.Pharm. 73(2) (2005

New metabolically stable fatty acid amide ligands of cannabinoid receptors: synthesis and receptor affinity studies

We investigated the structure-activity relationships for the interactions of fatty acid amide analogs of the endocannabinoid anandamide with human recombinant cannabinoid receptors. Thirty-five novel fatty acid amides were synthesized using five different types of acyl chains and 11 different aromatic amine 'heads.' Although none of the new compounds was a more potent ligand than anandamide, we identified three amine groups capable of improving the metabolic stability of arachidonoylamides and their CBI/CB2 selectivity ratio to over 20-fold, and several aromatic amines capable of improving the affinity of short chain or monosaturated fatty acids for cannabinoid CBI receptors. For the first time a tertiary amide of arachidonic acid was found to possess moderate affinity (K, = 300 nM) for cannabinoid CBI, but not CB2, receptors

Development of the first potent and specific inhibitors of endocannabinoid biosynthesis.

Enzyrnes for the biosynthesis and degradation of the endocannabinoid 2-arachidonoyl glycerol (2-AG) have been cloned and are the sn-1-selective-diacylglycerol lipases alfa and beta (DAGLalfa and beta) and the monoacylglycerol lipase (MAGL), respectively. Here, we used membranes from COS cells over-expressing recombinant human DAGLalfa to screen new synthetic substances as DAGLalfa inhibitors, and cytosolic fractions from wild-type COS cells to look for MAGL inhibitors. DAGLalfa and MAGL activities were assessed by using sn-1-[14C]-oleoyl-2- arachidonoyl-glycerol and 2-[3H]-arachidonoylglycerol as substrates, respectively. We screened known compounds as well as new phosphonate derivatives of oleic acid and fluoro-phosphinoyl esters of different length. Apart from the general lipase inhibitor tetrahydrolipstatin (orlistat) (IC50 60 nM), the most potent inhibitors of DAGLalfa were 0-3640 [octadec-9-enoic acid-l-(fluoro-methyl-phospboryloxymcthyl)-propylester] (lC50=500 nM). and 0-3841 [octadec-P-enoic acid l-methoxymethyl-2-(tluoro-methyl-phosphinoyloxy)-ethyl ester] (IC50= 160 nM). Apart from being almost inactive on MAGL, these two compounds showed high selectivity over rat liver triacylglycerol lipase, rat N-acylphosphatidyl-ethanolamine-selective phospholipase D (involved in anandamide biosynthesis), rat fatty acid amide hydrolase and human recombinant cannabinoid CB1 and CB2 receptors. Methylarachidonoyl-fluorophosphonate and the novel compound UP-l0l [O-ethyl-O-p-nitro-phenyl oleylphosphonate] inhibited both DAGLa and MAGL with similar potencies (JC50=O.8-0.1 and 3.7-3.2 11M, respectively). Thus, we report the first potent inhibitor of the biosynthesis of 2-AG that may be used as pharmacological tools to investigate the biological role of this cannabinoid